US3723411A

US3723411A - Beta-rhodomycin v

Info

Publication number: US3723411A
Application number: US00123882A
Authority: US
Inventors: H Brockmann; M Scheer
Original assignee: Bayer AG
Current assignee: Bayer AG
Priority date: 1970-03-18
Filing date: 1971-03-12
Publication date: 1973-03-27
Anticipated expiration: 1990-03-27
Also published as: IL36356A0; DE2012808A1; FR2085720B1; BE764472A; GB1292010A; FR2085720A1; NL7103590A

Abstract

An antibiotic designated Beta -rhodomycin V and corresponding to the formula WHEREIN R represents a tetraglycoside consisting of 2 mols rhodinose, 1 mol 2-desoxy-fucose and 1 mol rhodosamine is prepared by cultivating Streptomyces purpurascens, extracting the culture filtrate and mycelium in a neutral medium and obtaining the antibiotic from the extracts by known methods. The antibiotic is found to possess excellent activity against a broad class of organisms, especially mycoplasmas and gram-positive bacteria.

Description

United States Patent 1 Brockmann et al.

[54] BETA-RHODOMYCIN V [75] Inventors: Hans Brockmann, Goettingen; Martin Scheer, Wuppertal-Elberfeld, both of Germany [73] Assignee: Farbenfabriken Bayer Aktiengesellschait, Leverkusen, Germany 221 Filed: Mar. 12, 1971 21 Appl.No.: 123,882

[30] Foreign Application Priority Data Mar. 18, 1970 Germany ..P 20 12 808.9

[52] U.S. Cl. ..260/210 AB, 424/180,195/80 [51] Int. Cl ..C07c 47/18 [58] Field of Search ..260/210 AB, 210 R [56] References Cited OTHER PUBLICATIONS Bayer, Chem. Abst., Vol.53, 1959, p. 4663(b). Brockmann et al., Chem. Abst., Vol. 70, 1969, p. 97ll6y.

[ 3,723All 51 Mar. 27, 1973 Primary ExaminerLewis Gotte Assistant Examiner-Johnnie R. Brown AttorneyMcCarthy, DePaoli, OBrien & Price [57] ABSTRACT An antibiotic designated B-rhodomycin V and corresponding to the formula O-rliodosuminc OR OH O OH wherein R represents a tetraglycoside consisting of 2 mols rhodinose, 1 mol 2-desoxy-fucose and 1 mol rhodosamine is prepared by cultivating Streptomyces- 1 Claim, No Drawings BETA-RHODOMYCIN v BACKGROUND INVENTION 1 Field of the Invention The present invention relates in general to the field of antibiotics, more particularly to a new member of the class of rhodomycins.

2. Description of the Prior Art The rhodomycins are closely interrelated and are nitrogen-containing bases which usually occur as mixtures. They are red dyestuffs.

German Patent No. 851,398 is directed to a process for the production of antibiotically active substances, which is characterized in that streptomycetes, especially Streptomyces purpurascens novv spec., are allowed to grow under aerobic conditions on or in nutrient media known for the cultivation of streptomycetes and the rhodomycins are obtained from the resultant cultures by usual methods.

The subject matter of German Patent No. 913,813 is the preparation of a crystallized rhodomycin hydrochloride, designated therein as rhodomycin-A hydrochloride.

SUMMARY OF THE INVENTION The subject-matter of the present inventionis a new B-rhodomycin called B-rhodomycin V and correspondin to the formula R tetraglycoside from 2 mol rhodinose, 1 mol 2-desoxy-fucose and 1 mol rhodosamine.

This new rhodomycin is distinguished from the known rhodomycins in that it contains fivesugar radicals, whereas the known rhodomycins, now called B- rhodomycins I and B-rhodomycins II, only contain one and two sugar radicals, respectively [Tetrahedron Letters, 1969, pages 415 to 419].

According to the invention, the new rhodomycin is prepared by cultivating Streptomyces purpurascens, extracting the culture filtrate and vthe mycelium in a neutral medium, and obtaining B-rhodomycin V from the extracts in known manner.

The process according to the invention is thus distinguished from the known processes for the production of the known rhodomycins in that the extraction is here carried out in a neutral medium whereas in the earlier patents mentioned above the extraction is carried out in an acidic medium.

All species of streptomycetes are suitable starting materials for the process according to the invention. Streptomyces purpurascens Ist 313 is particularly suitable. Neutral extraction means in the present case an extraction at a pH value of 6.5 to 7.5.

fi-Rhodomycin V has an excellent chemotherapeutic, especially antibacterial activity, as can be seen from the following results of tests in vitro.

Testing of the minimal inhibition concentration was carried out for the gram-positive and gram-negative causative organisms in the KLEIN medium (meat extract, peptone, dextrose, pH 7.1) at an incubation temperature of 37C. for 24 hours, number of seeded germs l0 genns/ml; and for mycoplasmas in PPLO broth (PPLO basic substrate, dextrose, equine serum, pl-l7.6) at an incubation temperature of 37C. for 48 hours to 72 hours, number of seeded germs 10 germs/ml.

TABLE 1 Minimal Inhibition Concentration (MIC) (y/ml nutrient medium) of ,B-rhodomycin V of Type of germ strains MIC -y/ml Streptococci of different species 5 0.015 to 1.00 Staphylococcus aureus 4 0.050 to 0.12 Sarcinae 2 0.060 to 1.00 Corynebacterium pyogener 2 0.060 to 0.12 Pseudomona: aemginosa 2 E. coli 2 50 Klebsiella pneumom'ae l 25 Salmonella spec. 2 50 to 200 Parteurella multocida 2 6.25

Bordetella bronchiseptica 1 50 Alkaligenenesfaecalilr 1 25 Mycoplasmas avian mycoplasmas 4 0.001 to 0.002 bovine mycoplasmas 2 0.030 to 0.060 porcine mycoplasmas 2 0.001 to 0.002

The values of the Table show that B-rhodomycin V has an excellent and unusually wide-ranging activity, especially against mycoplasmas and gram-positive bacteria.

The excellent and wide-ranging activity permits of their use in human as well as veterinary medicine; they can be used for the prophylaxis of bacterial infections but also for the treatment of already existing bacterial infections.

As indications for the field of veterinary medicine, there may be mentioned, for example, infections with mycoplasmas and intestinal infections. For prophylaxis, the B-rhodomycin V can be used as admixture to feeding stuff.

EXAMPLE 1 A 10 liters preliminary culture of Streptomyces purpurascens lst 313 is incubated on a nutrient solution with 2 percent of soya flour and 2 percent mannitol submersed at 26C. for 24 hours, transferred to 350 liters of a nutrient solution of the same composition, and fermented for 96 hours at 260C. with stirring and aeration.

When the fermentation has been terminated, the redbrown culture broth is stirred with 20 kg kieselguhr and filtered. The culture filtrate and the mycelium are worked up separately. The mycelium is stirred three times with 60 liters of acetone each time, the combined orange-red acetone extracts are evaporated in a vacuum at 50C. to a volume of 30 liters, extracted three times with 15 liters of butanol each time, and the combined butanol extracts are concentrated in a vacuum at 50C. to volume of about 3 liters. The remaining red-brown oil is taken up with 10 liters chloroform (solution A).

The culture filtrate (350 liters, pH 7.0 to 7.5) is extracied with 150 liters butanol and the deep-red butanol phase is concentrated in a vacuum at 50C. until there remain about 3 liters of a deep-red oil which is taken up with 10 liters chloroform (solution B).

The combined chloroform solutions A and B are distributed over six 100 X 10 cm columns of carboxymethyl cellulose (deposited with 2N hydrochloric acid; washed out (1) with water until the reaction is neutral, (2) with acetone and (3) with chloroform). Rinsing with 5 to liters chloroform per column brings fats, anthracyclinones and non-basic anthracyclines into the filtrate.

The portion remaining on the column is eluted with 10 percent aqueous potassium hydrogen phosphate, and the deep-red eluate (5 liters per column) which has been adjusted to pH 8.0 with dilute ammonia is extracted three times with three liters of chloroform each time. The combined chloroform phases are evaporates to dryness in a vacuum, the residue is taken up with a little chloroform, and the red B-rhodomycin V is precipitated with petroleum ether (40 to 50C.). Yield 150 to 170 mg per liter of culture broth.

2 g of the B-rhodomycin V preparation obtained as described above are chromatographed through a 100 X 4 cm cellulose column in a system butanol-0.7 m phosphate buffer pH 5.8 1:1 The main zone (migrating fastest) is brought into the filtrate by rinsing with r phase, dilute ammonia is then added to the filt; zte until the pH is 8.0, the mixture is shaken with chloroform, and the chloroform phase, dried over sodium sulphate, is evaporated to dryness in a vacuum. The

B-rhodomycin V remaining behind as a red lacquer is taken up with a little chloroform and precipitated in the form of a red powder or also of red needles at boiling temperature with cyclohexane. Melting point 173 to 175C. (decomposition); [a 1 m 40 i 2C, 0.2 in chloroform. C H N O 1059.2)

Calc.: C 61.14; H 7.36; N 2.61

found: C 61.30; H 7.40; N 2.65 dried in a high vacuum at C. for 15 hours.

IR (KBr compact) 3450, 2910, 1730, 1595/cm.

The B-rhodomycin V is readily soluble in methanol, butanol, acetone, chloroform, 0.7 m phosphate buffer pH 5.8, dilute acids; it is sparingly soluble in benzene, very sparingly soluble in water and virtually insoluble in cyclohexane.

What is claimed is:

1. B-Rhodomycin V of the formula on o CHJCI'IQ I 1 My 1 on on 3 on in which R stands for a tetraglycoside radical consisting of 2 mol rhodinose, 1 mol 2-desoxy-fucose and 1 mol rhodosamine.